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PMID: 2566911 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Sequences downstream of AAUAAA signals affect pre-mRNA cleavage and polyadenylation in vitro both directly and indirectly.

Molecular and cellular biology ·Vol. 9 ·No. 4 ·1989-04-00 ·Pages 1759-71

Ryner LC, Takagaki Y, Manley JL

Abstract

To investigate the role of sequences lying downstream of the conserved AAUAAA hexanucleotide in pre-mRNA cleavage and polyadenylation, deletions or substitutions were constructed in polyadenylation signals from simian virus 40 and adenovirus, and their effects were assayed in both crude and fractionated HeLa cell nuclear extracts. As expected, these sequences influenced the efficiency of both cleavage and polyadenylation as well as the accuracy of the cleavage reaction. Sequences near or upstream of the actual site of poly(A) addition appeared to specify a unique cleavage site, since their deletion resulted, in some cases, in heterogeneous cleavage. Furthermore, the sequences that allowed the simian virus 40 late pre-RNA to be cleaved preferentially by partially purified cleavage activity were also those at the cleavage site itself. Interestingly, sequences downstream of the cleavage site interacted with factors not directly involved in catalyzing cleavage and polyadenylation, since the effects of deletions were substantially diminished when partially purified components were used in assays. In addition, these sequences contained elements that could affect 3'-end formation both positively and negatively.

MeSH Terms
Adenoviridae/genetics,metabolism Base Sequence Binding Sites Chromosome Deletion HeLa Cells/metabolism Humans Molecular Sequence Data Poly A/genetics,metabolism RNA Precursors/genetics,metabolism RNA, Messenger/genetics,metabolism RNA, Viral/genetics,metabolism Simian virus 40/genetics,metabolism
Chemicals
RNA Precursors RNA, Messenger RNA, Viral Poly A
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Ryner L C
Department of Biological Sciences, Columbia University, New York, New York 10027.
Takagaki Y
Manley J L
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.9.4.1759-1771.1989
Published
1989-04-00
Pages
1759-71
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC362595
Subset
IM
Grants
NIGMS NIH HHS · GM 28983 · United States
External Links
PubMed source DOI: 10.1128/mcb.9.4.1759-1771.1989 PMC Full Text View journal details
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